microRNA miR-144 modulates oxidative stress tolerance and associates with anemia severity in sickle cell disease.

Although individuals with homozygous sickle cell disease (HbSS) share the same genetic mutation, the severity and manifestations of this disease are extremely heterogeneous. We have previously shown that the microRNA expression in normal and HbSS erythrocytes exhibit dramatic differences. In this study, we identify a subset of HbSS patients with higher erythrocytic miR-144 expression and more severe anemia. HbSS erythrocytes are known to have reduced tolerance for oxidative stress, yet the basis for this phenotype remains unknown. This study reveals that miR-144 directly regulates nuclear factor-erythroid 2-related factor 2, a central regulator of cellular response to oxidative stress, and modulates the oxidative stress response in K562 cell line and primary erythroid progenitor cells. We further demonstrate that increased miR-144 is associated with reduced NRF2 levels in HbSS reticulocytes and with decreased glutathione regeneration and attenuated antioxidant capacity in HbSS erythrocytes, thereby providing a possible mechanism for the reduced oxidative stress tolerance and increased anemia severity seen in HbSS patients. Taken together, our findings suggest that erythroid microRNAs can serve as genetic modifiers of HbS-related anemia and can provide novel insights into the clinical heterogeneity and pathobiology of sickle cell disease.

[1]  E. Miska.  Faculty Opinions recommendation of The miR-144/451 locus is required for erythroid homeostasis. , 2010 .

[2]  Michel Sadelain,et al.  A Genetic Strategy for Single and Combinatorial Analysis of miRNA Function in Mammalian Hematopoietic Stem Cells , 2009, Stem cells.

[3]  H. Grosshans,et al.  Active turnover modulates mature microRNA activity in Caenorhabditis elegans , 2009, Nature.

[4]  T. Katoh,et al.  Selective stabilization of mammalian microRNAs by 3' adenylation mediated by the cytoplasmic poly(A) polymerase GLD-2. , 2009, Genes & development.

[5]  S. Biswal,et al.  Nrf2-regulated glutathione recycling independent of biosynthesis is critical for cell survival during oxidative stress. , 2009, Free radical biology & medicine.

[6]  Saijuan Chen,et al.  Mir-144 selectively regulates embryonic alpha-hemoglobin synthesis during primitive erythropoiesis. , 2009, Blood.

[7]  V. Ambros,et al.  The evolution of our thinking about microRNAs , 2008, Nature Medicine.

[8]  Xuemei Chen,et al.  Degradation of microRNAs by a Family of Exoribonucleases in Arabidopsis , 2008, Science.

[9]  Yulei N. Wang,et al.  The Genomic Analysis of Erythrocyte microRNA Expression in Sickle Cell Diseases , 2008, PloS one.

[10]  Xiaowu Gai,et al.  A GATA-1-regulated microRNA locus essential for erythropoiesis , 2008, Proceedings of the National Academy of Sciences.

[11]  Stijn van Dongen,et al.  miRBase: tools for microRNA genomics , 2007, Nucleic Acids Res..

[12]  M. Socolovsky.  Molecular insights into stress erythropoiesis , 2007, Current opinion in hematology.

[13]  Shyam Biswal,et al.  Cell survival responses to environmental stresses via the Keap1-Nrf2-ARE pathway. , 2007, Annual review of pharmacology and toxicology.

[14]  S. Bonovas,et al.  Redox imbalance, macrocytosis, and RBC homeostasis. , 2006, Antioxidants & redox signaling.

[15]  E. Lam.  Faculty Opinions recommendation of Posttranscriptional induction of two Cu/Zn superoxide dismutase genes in Arabidopsis is mediated by downregulation of miR398 and important for oxidative stress tolerance. , 2006 .

[16]  W. Filipowicz,et al.  Relief of microRNA-Mediated Translational Repression in Human Cells Subjected to Stress , 2006, Cell.

[17]  G. Longmore.  A unique role for Stat5 in recovery from acute anemia. , 2006, The Journal of clinical investigation.

[18]  E. Fibach,et al.  Red blood cells, platelets and polymorphonuclear neutrophils of patients with sickle cell disease exhibit oxidative stress that can be ameliorated by antioxidants , 2006, British journal of haematology.

[19]  Mark T Gladwin,et al.  Lactate dehydrogenase as a biomarker of hemolysis-associated nitric oxide resistance, priapism, leg ulceration, pulmonary hypertension, and death in patients with sickle cell disease. , 2005, Blood.

[20]  Paola Sebastiani,et al.  Genetic dissection and prognostic modeling of overt stroke in sickle cell anemia , 2005, Nature Genetics.

[21]  Jiang Li,et al.  Stabilization of Nrf2 by tBHQ confers protection against oxidative stress-induced cell death in human neural stem cells. , 2005, Toxicological sciences : an official journal of the Society of Toxicology.

[22]  C. Burge,et al.  Conserved Seed Pairing, Often Flanked by Adenosines, Indicates that Thousands of Human Genes are MicroRNA Targets , 2005, Cell.

[23]  Masayuki Yamamoto,et al.  Nrf2-Keap1 defines a physiologically important stress response mechanism. , 2004, Trends in molecular medicine.

[24]  Y. Kan,et al.  Targeted disruption of Nrf2 causes regenerative immune-mediated hemolytic anemia. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[25]  W. Klitz,et al.  Gene interactions and stroke risk in children with sickle cell anemia. , 2004, Blood.

[26]  C. Brugnara,et al.  Pathophysiological-based approaches to treatment of sickle cell disease. , 2003, Annual review of medicine.

[27]  S. Orkin,et al.  Essential role for the peroxiredoxin Prdx1 in erythrocyte antioxidant defence and tumour suppression , 2003, Nature.

[28]  Y. Kan,et al.  Identification of the NF-E2-related Factor-2-dependent Genes Conferring Protection against Oxidative Stress in Primary Cortical Astrocytes Using Oligonucleotide Microarray Analysis* , 2003, The Journal of Biological Chemistry.

[29]  W. Klitz,et al.  Distinct HLA associations by stroke subtype in children with sickle cell anemia. , 2003, Blood.

[30]  R. Tibshirani,et al.  Diagnosis of multiple cancer types by shrunken centroids of gene expression , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[31]  A. Calzolari,et al.  Accentuated response to phenylhydrazine and erythropoietin in mice genetically impaired for their GATA-1 expression (GATA-1(low) mice). , 2001, Blood.

[32]  S. Mahajan,et al.  Glutathione protects chemokine-scavenging and antioxidative defense functions in human RBCs. , 2001, American journal of physiology. Cell physiology.

[33]  J. Chan,et al.  Impaired expression of glutathione synthetic enzyme genes in mice with targeted deletion of the Nrf2 basic-leucine zipper protein. , 2000, Biochimica et biophysica acta.

[34]  Christian A. Rees,et al.  Molecular portraits of human breast tumours , 2000, Nature.

[35]  Ken Itoh,et al.  Transcription Factor Nrf2 Coordinately Regulates a Group of Oxidative Stress-inducible Genes in Macrophages* , 2000, The Journal of Biological Chemistry.

[36]  Ash A. Alizadeh,et al.  Distinct types of diffuse large B-cell lymphoma identified by gene expression profiling , 2000, Nature.

[37]  K. Itoh,et al.  An Nrf2/small Maf heterodimer mediates the induction of phase II detoxifying enzyme genes through antioxidant response elements. , 1997, Biochemical and biophysical research communications.

[38]  V. Tatum,et al.  Antioxidant status and susceptibility of sickle erythrocytes to oxidative and osmotic stress. , 1996, Free radical research.

[39]  C. Pegelow,et al.  Clinical events in the first decade in a cohort of infants with sickle cell disease. Cooperative Study of Sickle Cell Disease. , 1995, Blood.

[40]  G. Rodgers,et al.  Mortality in sickle cell disease. , 1994 .

[41]  O. Castro,et al.  The acute chest syndrome in sickle cell disease: incidence and risk factors. The Cooperative Study of Sickle Cell Disease. , 1994, Blood.

[42]  O. Platt,et al.  Mortality in sickle cell disease. Life expectancy and risk factors for early death. , 1994, The New England journal of medicine.

[43]  B. Fanburg,et al.  Regulation of cellular glutathione. , 1989, The American journal of physiology.

[44]  J W Eaton,et al.  Pathobiology of heme interaction with the erythrocyte membrane. , 1989, Seminars in hematology.

[45]  A. Prasad,et al.  Altered amount and activity of superoxide dismutase in sickle cell anemia , 1988, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[46]  R. Hebbel,et al.  Abnormal redox status of membrane-protein thiols in sickle erythrocytes. , 1985, The Journal of clinical investigation.

[47]  J. Eaton,et al.  Spontaneous oxygen radical generation by sickle erythrocytes. , 1982, The Journal of clinical investigation.

[48]  B. Lubin,et al.  Abnormal vitamin E and glutathione peroxidase levels in sickle cell anemia: evidence for increased susceptibility to lipid peroxidation in vivo. , 1979, The Journal of laboratory and clinical medicine.

[49]  B. Ames,et al.  Erythrocyte glutamine depletion, altered redox environment, and pulmonary hypertension in sickle cell disease , 2009 .